Isomer of cholesterol and process for producing the same



Pa'tented June 25, 1940 ISOMER F CHOLESTEROL AND rRooEss FOR raonncmarna SAME Everett S. Wallis, Princeton, and Erhard Fernholz, Elizabeth, N. .L, assignors to Merck 8; Co; Inc., Railway, N. J., a corporation-oi New Jersey No Drawing. Application January 16, 1937,

.. Serial N0. 120,880

'2 Claims. (Cl. 260-397) 1' This invention relates to new compounds called i-cholesteryl acetate and i-cholesterol, (i indigiating isomer) and to processes for their. produc- 3 These new compounds were prepared by us during the course of our investigations of new processes for the preparation, of the epimeric modifications of certain unsaturated sterols.

The epimerization of the hydroxyl group of cholesterol, and other unsaturated sterols, has assumed considerable importance since Ruzicka et al. lHelv. Chim. Acta. 17 1395 (1934)] reported their findings to the eflect that the male hormone, androsterone, is a derivative of epi-dihy- 5 drocholesterol, and-that its, physiological activity is much greater than that of the corresponding epimer, iso-androsterone, obtained from p-cholestanol.

Our method of approach to the preparation of epi-cholesterol, was based .upon certain experiments reported by Phillips (J. Chem. Soc. 123 44 (1925)), carried out on alcohols containing only one asymmetric carbon atom, and it involved an investigation of the action of potassium acetate on cholesteryl-p-toluene sulfonate. 'However, in

order to avoid the complication ofthe formation 0! ethers, and since we had observed that, in

- acetic acid solution, the normal cholesteryl acetate is formed, our reaction was carried out with an- 0 hydrous potassium acetate in acetic anhydride solution. 'Very unexpectedly, a new, hitherto unknown, acetate was thus obtained, which is iso-.

meric with cholesteryl acetate, and which we. call i-cholesteryl acetate.

This new compound is strongly dextro-rotatory,

. [4% and melts-at about 73C." On hydrolysis, it yields i-cholesterol. It crystallizes from alcohol-in long needles which melt at room temperature. The substance resolidifies and passes into-another crystalline form which melts at "24-75" C. The hydrolysis product, i-cholesterol, is. also strongly dextro-rotatory -[a],,'=+23;9

It is not precipitated by digitonin, Reacetylation yields the abovei-oholesteryli acetate.

Hydrogenation of the new acetate is most satissolution' of potassium carbonate. The acetic an-z factorily accomplishedby reacting upon it with the Adams platinum oxide catalyst. When this platinum oxide is reduced in the presence of the. i -cholesteryl acetate, rapid hydrogenation takes place and dihyd'rocholesteryl acetate is formed ingood. yield. Palladium black was found to be ineffective for this purpose, while only a very slow reaction is obtained with platinum black.

Our new acetate, i-cholesteryl acetate, does not react with perbenzoic acid. It doesnot de- 10 coloriz e readily a solutionof bromine in carbon tetrachloride. 'The Liebermann reaction is quite strong. This inertness toward bromine and perbenzoic acid, and its relative stability toward'catl allytic hydrogenation suggest that a double bond, 15 as such, is not present in i-cholesteryl acetate and i-oholesterol. v

We have found that i-cholesterol may be easily oxidized withchromic acid in acetic acid solution. From the products formed, an oxime has 20 been isolated, in 20% yield. This indicates that i-cholesterol is a secondary alcohol.

Preparation of i-cholesteryl acetate About -100 gms. of anhydrous potassium acetate 25 aredissolved in 1500 cc. of boiling pure acetic anhydride. The solution is cooled to C.

-whereupon potassium acetate crystallizes in a finelyvdivided state. About 35 gms. of choles-' 30 teryl-p-toluene sulfonate are added. The flask is then placed on the steam bath, and stirred v for approximately 36 hours, during which time:

the temperature inside the flask is varied from '70 to C. The dark brown mixture is poured into cold water; 800 cc. of ether are added, and the ether layer iszwa'shed repeatedly with a cold aqueous hydridealso may be removed by distillation un-' der diminished. pressure. After complete fremovalofth'e acetic anhydr'ide and acetic acid,'the

etlier layer is dried with anhydrous sodium s'uli ate decolorized with animal charcoal and the 45 ether removed by distillation;

An pay product is obtained. This material is dissolved in alcohol, and small amounts of ether are added. Crystallization may be carried out in such a manner as to obtain four crops of crystals:

1st crop wt. 3.6 gms. M. P. 71.5" C.

2nd crop wt. 0.5 gm. M. P. IO-71 C. 3rd crop wt. 4.7 ms. M. P. fit-66 C. 4th crop wt. 7.2 gms. M. P. 59-62" 0.

Several recrystallizations of the 3rd and 4th fractions give 0.4 gm. of material having a melting point of 71 C. and 7.5 ms. of a material melting at 67-68 C. i

The i-cholesteryl acetate may be freed from cholesteryl acetate which occurs with it in the following manner:

About 7 /2 gms. of i-cholesteryl acetate (is. P.

67-68 C.) are dissolved in 100 cc. of alcohol, and treated with 8 guns. of potassium hydroxide. The solution is refluxed for one-half hour on the water bath, then poured into water and extracted with ether. The dried ether solution is evaporated to dryness and taken up in a small amount of 90% alcohol. A solution of 6 gms. of digitoninin 500 cc.

of 90% alcohol is then added. The contents are allowed to stand until they reach room temperature. The digitonide is filtered and dried.

The filtrate is evaporated to dryness and digested with ether. The ether solution is distilled and the residue is dissolved in alcohol. On cooling in an ice-salt mixture, i-cholesterol crystallizes. The crystals are filtered rapidly, and placed in a vacuum desiccator. They melt and resolidify. Weight of product, 3.5 gms.; melting point, I l-75 C.

About 22.6 mg. of this material dissolved in 2 cc. of chloroform solution gave an=+027 (100 mm. tube) About 1.5 ms. of i-cholesterol is heated with 10 cc. of acetic anhydride for about one hour on the water bath. on cooling, a crystalline product separates. Recrystallization from alcohol gives an acetate which melts at 73 C.

About 25.1 mg. in 2 cc. chloroform solution gave ap=+0.6.

Oxidation of i-cholesterol with chromic acid About ninety-three hundredths of a gram of i-cholesteroi is dissolved in 30 cc. of specially purified acetic acid (KMNOt method). A solution of 0.64 gm. of chromic acid in cc. of acetic acid is added drop-by-drop. The solution becomes temporarily cloudy, and the temperature rises slightly. The mixture is kept at room temperature for about 12 hours, and is then extracted with ether. The ether is shaken with an aqueous solutionof 2N sodium hydroxide to remove acidic material. This acidic material cannot be cmtallized. The neutral part crystallizes as beholesterone. It forms an oxime.

Prepcratiim o! the crime About thirty-three hundredths of-a gram of the crude ketone, l-cholesterone, described above. is boiledfor two hours with 0.3 mod hydraulamlne hydrochloride and 0.5 gm. of sodium scetate in cc. of ethyl alcohol. Thesoluticnis diluted with water. and the finely divided white material is filtered. tion from dilute alcohol gives i-cholesterone crime in the form or small leaflets which. when thoromhly d1! melt at 148-144 C.

The new compounds have the structural formula where B. may be selected from the group consisting of an acetate radical, hydroxyl, oxygen, and the oxime radical.

We claim as our invention:

1. The product of reaction between cholesterylp-toluene suifonate and anhydrous potassium acetate in acetic anhydride.

2. The hydrolysis product of the product covered by claim 1.

3. The process for the production of i-cholesteryl acetate having the following formula:

OH: OH: (EH-CEOHrCHsO I 03300 which comprises reacting upon anhydrous potassium acetate in acetic anhydride with cholesteryl-p-toluene sulfonate.

4. The process for the production of i-choleaterol having the following formula:

OH: OH:

BIb-OHsOHaOHrO which comprises reacting upon anhydrous potassium acetate in acetic anhydride with cholesteryl-p-toluene sulfonate, and hydrolyzing the i-cholesteryl acetate thus produced to obtain i-cholesterol.

s. A product of th'e'formula on. c tn-cal'onlomc 7 a,aos,44e' I 3 where B may be selected from the group con- 7. l-Cholesterol of the following tormula: sisting of an acetate radical, hydroxyl, oxygen. and the oxime radical,

6. i-cholesteryl acetate 0! the following for- 6 mulaf cm om eri-omomomo a I Om om om ea-omomomofi OBI mvmm'r s. WALLIS} mmmn mom. 

